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First published online 13 March 2007
doi: 10.1242/jcs.03424
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Research Article |
B signaling by the actin cytoskeleton
1 Laboratory of Virology and Immunology, CBIG-GIGA, University of Liège, Liège, Belgium
2 Laboratory of Microbiology, Institute for Microbiological Research J.-M. Wiame, University of Brussels, Brussels, Belgium
3 GSF-Institut für Molekulare Immunologie, Marchioninistr. 25, 81377 München, Germany
4 Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur and INSERM, U786, Paris 75724 CEDEX 15, France
* Author for correspondence (e-mail: S.Legrand{at}ulg.ac.be)
Accepted 6 February 2007
Actin disruption by CytochalasinD (CytD) and LatrunculinB (LatB) induced NF-
B activation in myelomonocytic and intestinal epithelial cells. In an attempt to elucidate the mechanism by which actin disruption induced IKK activation, we studied the human Nod2 protein, which was able to induce NF-
B activation and whose expression was restricted to myelomonocytic and intestinal epithelial cells. Nod2 is thought to play key roles in pathogen defence through sensing bacteria and generating an inflammatory immune response. We showed that actin disruption by CytD significantly and specifically increased Nod2-mediated NF-
B signaling. Nod2 was fully partitioned in the Triton-X-100-insoluble fraction but translocated into the soluble fraction after CytD treatment, demonstrating that the presence of Nod2 in the detergent-insoluble pellet was specific to actin cytoskeleton. Confocal analysis also revealed a Nod2 colocalization with membrane-associated F-actin. Colocalization and co-immunoprecipitation assays with endogenous Rac1 have shown that Nod2 associated with activated Rac1 in membrane ruffles through both its N-terminal caspase recruitment domains (CARD) and C-terminal leucine-rich repeats (LRRs). Membrane ruffle disruption by a Rac1 dominant negative form primed Nod2-dependent NF-
B signaling. The recruitment of Nod2 in Rac-induced dynamic cytoskeletal structures could be a strategy to both repress the Nod2-dependent NF-
B signaling in unstimulated cells and rapidly mobilize Nod2 during bacterial infection.
Key words: Actin cytoskeleton, NF-
B, Nod2, Rac1
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